ABSTRACT
The application of molecular techniques specically developed for clinical and research settings long predate the Human Genome Project. Molecular pathological methods were used to explicate the genetic basis of many diseases, and these innovations eventually contributed to the discipline of molecular diagnostics. Insights gained from diagnostics have led to improvements in genetic and molecular approaches to the classication of human cancers. This has developed, through multiparallelized approaches, into the validation of predictive biomarkers for medical response, disease progression, and the susceptibility of individual patients to develop malignancies. Today, molecular diagnostics continues to develop rapidly as diagnosis becomes inextricably linked to therapy.1,2
11.1 Introduction .......................................................................................................................... 159 11.2 Implications of Mathematical Modeling .............................................................................. 160 11.3 Dened Sample Handling and Preparation: A Prerequisite for Meaningful
Quantication ..................................................................................................................... 160 11.4 Sample Processing Solutions Provided by Microuidic Technologies................................. 161 11.5 Problems Associated with Measuring Heterogeneous Populations ..................................... 163 11.6 Future Perspective ................................................................................................................ 163 11.7 Methods ................................................................................................................................ 164
11.7.1 Droplet-Based PCR ................................................................................................... 164 11.7.2 Fabrication and Machining ....................................................................................... 164 11.7.3 Optimization of Flow Rates and Droplet Formation ................................................ 165 11.7.4 Heating System ......................................................................................................... 165 11.7.5 Analysis of PCR Products ........................................................................................ 165
11.8 Results ................................................................................................................................... 165 11.8.1 Droplet-Based PCR ................................................................................................... 165 11.8.2 Impact of Oil Flow Rates on the Droplet Production and Exposure Times ............. 166 11.8.3 Impact of Aqueous Flow Rates on the Droplet Production ...................................... 167 11.8.4 Analysis of PCR Products ........................................................................................ 167
11.9 Discussion ............................................................................................................................. 170 References ...................................................................................................................................... 170
The overall resolution of molecular analysis is improving, particularly as the eld reaches a crossroad whereby discrete genetic or other biomarker analysis subsides in the wake of gene-network analyses, enabled through holistic, systems-based, biological modeling of disease-causing pathways. This improvement also comes with a deepening and rmer comprehension of disease mechanisms, the benign term “biomarkers” conceding to more functionally related molecular descriptors.3
